Heterologna ekspresija gena TaMAB2 u transgenim biljkama Arabidopsis thaliana

master's thesis

• Title (english): Heterologous expression of TaMAB2 gene in transgenic Arabidopsis thaliana
• Author: Marija Klasić
• Mentor: Nataša Bauer (mentor)
• Committee member: Nataša Bauer (predsjednik povjerenstva)
• Committee member: Gordana Lacković-Venturin (član povjerenstva)
• Committee member: Željka Vidaković-Cifrek (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2012-02-15, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Proteini MAB sadrţe dvije funkcionalno bitne interakcijske domene: MATH i BTB. Do sada je pokazano da su oni dio sustava eliminacije proteina koji stanici više nisu potrebni. Neki od istraženih proteina MAB (CeMEL-26, HsSPOP) dio su multiproteinskih kompleksa kulin 3 E3 ligaza i sluţe za prepoznavanje supstrata koje treba razgraditi. Tijekom razvoja pšenice ekspresija gena TaMAB2 prisutna je samo u oplođenoj jajnoj stanici, u zigoti te u dvostaničnom embriju. U ovom radu napravljena je heterologna ekspresija gena TaMAB2 iz pšenice u uročnjaku. Utvrđeno je da transgenične biljke proizvode cjeloviti protein TaMAB2 koji se u stanicama nakuplja. Sam protein TaMAB2 je ubikvitiniran, te vjerojatno stvara homo- i heterodimere. Rast i razvoj transgeničnih biljaka je značajno fenotipski promijenjen u odnosu na biljke divljeg tipa ali im fototropizam i gravitropizam nisu poremećeni. Analizom unutarstanične lokalizacije proteina TaMAB2 u stanicama BY-2 i pokožici luka utvrđeno je nakupljanje velikih agregata oko jezgrine ovojnice i u blizini stanične membrane. Ovi rezultati ukazuju da bi protein TaMAB2 mogao imati bitnu ulogu u eliminaciji proteina i uspostavljanju polarnosti tijekom rane embriogeneze pšenice.

Abstract (english)

MAB proteins contain two functionally important interaction domains: MATH and BTB. It is known that MAB proteins function as a part of system by which cells eliminate unwanted proteins. Some MAB proteins (CeMEL-26, HsSPOP) are part of cullin 3 E3 ligase enzymatic complex and function as substrate adaptors. In wheat, TaMAB2 gene is specifically expressed only in fertilized egg cell, zygote and two-cell embryo. In this research heterologous expression of TaMAB2 gene in Arabidopsis was induced. High amount of TaMAB2 protein is accumulated inside the cells of transgenic plants. In transgenic plants TaMAB2 is ubiquitinated and probably assembled in homo- and heterodimers. Growth and development of transgenic plants are significantly changed regarding to wild type plants, but transgenic plants have normal phototropism and gravitropism. In BY-2 cells and onion epidermal cells TaMAB2 protein form aggregates that localizes unilaterally around nuclear envelope and near cell membrane. This results indicate on the important role of TaMAB2 in protein elimination and establishment of cell polarity during early embryogenesis of wheat.

• Keywords: proteinske domene MATH i BTB; razgradnja proteina; kulin 3 E3 ligaze; fenotipske promjene; fototropizam; gravitropizam; unutarstanična lokalizacija; transgenični uročnjak Arabidopsis thaliana
• Keywords (english): MATH and BTB protein domains; protein degradation; CUL3 E3 ligase; fenotypical changes; fototropism; gravitropism; subcellular localization; transgenic Arabidopsis thaliana
• Language: croatian
• URN:NBN: urn:nbn:hr:217:563581
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-02 08:42:40